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Peripheral artery disease is a progressive disease. Primary ischemic leg symptoms are muscle fatigue, discomfort or pain during ambulation, known as intermittent claudication. The most severe manifestation of peripheral artery disease is critical limb ischemia (CLI). The long-term safety of gene therapy in peripheral artery disease remains unclear. This four center peripheral artery disease registry was designed to evaluate the long-term safety of the intramuscular non-viral fibroblast growth factor-1 (NV1FGF), a plasmid-based angiogenic gene for local expression of fibroblast growth factor-1 versus placebo in patients with peripheral artery disease who had been included in five different phase I and II trials. Here we report a 3-year follow-up in patients suffering from CLI or intermittent claudication. There were 93 evaluable patients, 72 of them in Fontaine stage IV (47 NV1FGF versus 25 placebo) and 21 patients in Fontaine stage IIb peripheral artery disease (15 NV1FGF versus 6 placebo). Safety parameters included rates of non-fatal myocardial infarction (MI), stroke, death, cancer, retinopathy and renal dysfunction. At 3 years, in 93 patients included this registry, there was no increase in retinopathy or renal dysfunction associated with delivery of this angiogenic factor. There was also no difference in the number of strokes, MI or deaths, respectively, for NV1FGF versus placebo. In the CLI group, new cancer occurred in two patients in the NV1FGF group. Conclusions that can be drawn from this relatively small patient group are limited because of the number of patients followed and can only be restricted to safety. Yet, data presented may be valuable concerning rates in cancer, retinopathy, MI or strokes following angiogenesis gene therapy in the absence of any long-term data in angiogenesis gene therapy. It may take several years until data from larger patient populations will become available.

Restenosis remains the major limitation of percutaneous transluminal coronary angioplasty. Restenosis after balloon angioplasty is due to vascular remodeling and neointimal hyperplasia. In spite of encouraging results in animal models, most of the pharmacological trials of prevention of restenosis in humans have produced negative results. This has prompted interest in the potential role of locally delivered drugs and various balloon catheter systems that are now available to achieve local delivery of therapeutic agents at the site of arterial injury. In 1997, implantation of a coronary stent in conjunction with balloon angioplasty is performed in an increasing number of patients. Randomized studies have shown that coronary stenting may reduce the risk of restenosis. In addition, restenosis after coronary stenting is mainly due to neointimal hyperplasia. Restenosis within coronary stents might thus be much more sensitive to therapies designed to inhibit neointimal hyperplasia than restenosis after balloon angioplasty. Thus, the future prevention of restenosis might well be the combination of a mechanical device that produces the widest possible lumen and prevents vessel constriction with a pharmacologic approach to inhibit the proliferative process. (Trends Cardiovasc Med 1997;7:90-94). © 1997, Elsevier Science Inc.

Objectives. To evaluate clinical events in a specifically selected cohort of patients with obstructive coronary artery disease (CAD), using a new generation thin-strut bare cobalt-chromium coronary stent.Methods. Patients with single- or multi-vessel, stable or unstable CAD eligible for percutaneous implantation of at least one bare cobalt-chromium stent were evaluated in a single-centre registry. Prospective pre-specified criteria for bare cobalt-chromium stent implantation in our centre were: any acute ST-elevation myocardial infarction (MI), otherwise 1) de novo coronary lesion, and 2) lesion length20 mm, and 3) reference vessel diameter2.6 mm, and 4) no diabetes, unless reference vessel diameter3.5 mm. Endpoints, retrospectively collected, were death, MI and clinically driven target-lesion revascularisation (TLR) and target-vessel revascularisation (TVR) after 12 months.Results. Between September 2005 and June 2007, 712 patients (48.7% one-vessel, 29.9% two-vessel, 20% three-vessel and 1.4% left main disease; 7.9% diabetics) were treated with 800 bare cobalt-chromium stents, for stable angina (40.9%), unstable angina (20.9%) or acute ST-elevation MI (38.2%). The procedural success rate was 99.3%. Peri-procedural MI rate was 2.2% in the semi-elective group. At 12 months there were 17 deaths (2.4%), of which nine non-cardiac, 20 (2.8%) MI, 19 (2.7%) TLR and 29 (4.1%) TVR. Early and late definite stent thrombosis occurred in four (0.6%) and three (0.4%) patients, respectively.Conclusion. A strategy aimed at minimising drug-eluting stent use and combining a pre-specified simple selection process with the use of a new thin-strut bare cobalt-chromium stent is safe and effective at one-year clinical follow-up. (Neth Heart J 2010;18:486-92.).

Summary. Within the last 6 years, it has been demonstrated that drug-eluting stents (DES) reduce significantly angiographic and clinical restenosis after percutaneous coronary interventions. These results are consistent across several clinical randomized controlled trials comparing these new devices with bare metallic stents (BMS), which themselves have already markedly improved the results obtained with balloon angioplasty in the early days of this method of myocardial revascularization. Nevertheless, some concerns have been raised regarding a delayed endothelialization of the coated prostheses leading to late stent thrombosis occurring mainly when antiplatelet therapy is discontinued in the follow-up. The most recent data show that, in comparison with BMS, there is a small excess of late (> 1 year) stent thrombosis but this is not associated with an increased risk of death or myocardial infarction or all cause mortality. These concerns do not outweigh the strong benefits of DES in preventing restenosis but require a number of measures concerning a longer dual antiplatelet treatment (than initially expected), to control patient treatment compliance and to provide a complete education of patients and physicians. Future devices dealing with the two issues (antiproliferative properties with rapid controlled endothelialization preventing thrombosis) would be the next major advance in this rapidly evolving field.

Objectives: To assess non-invasively the acute effects of cardiac resynchronisation therapy (CRT) on functional mitral regurgitation (MR) at rest and during dynamic exercise. Methods: 21 patients with left ventricular (LV) systolic dysfunction and functional MR at rest, treated with CRT, were studied. Each patient performed a symptom-limited maximal exercise with continuous two dimensional Doppler echocardiography twice. The first exercise was performed with CRT; the second exercise was performed without CRT. Mitral regurgitant flow volume (RV), effective regurgitant orifice area (ERO) and LV dP/dt were measured at rest and at peak exercise. Results: CRT mildly reduced resting mitral ERO (mean 8 (SEM 2) v 11 (2) mm 2 without CRT, p = 0.02) and RV (13 (3) v 18 (3) ml without CRT, p = 0.03). CRT attenuated the spontaneous increase in mitral ERO and RV during exercise (1 (1) v 9 (2) mm 2 , p = 0.004 and 1 (1) v 8 (2) ml, p = 0.004, respectively). CRT also significantly increased exercise-induced changes in LV dP/dt (140 (46) v 479 (112) mm Hg/s, p Conclusion: Attenuation of functional MR, induced by an increase in LV contractility during dynamic exercise, may contribute to the beneficial clinical outcome of CRT in patients with chronic heart failure and LV asynchrony.

Interventional procedures associated with acute coronary syndromes or performed on saphenous bypass grafts frequently lead to embolic complications, resulting in no-reflow phenomenon, side-branch occlusion, or peri-procedural infarction. The RESCUE thrombo-aspiration system was used in 19 percutaneous coronary interventions. After initial use of the aspiration device, 81% of procedures were followed by stent deployment. TIMI flow 2 or higher was present in 42% at the beginning of the procedure and in 95% at the end. In-hospital MACE rate was 4.76%. This relatively user-friendly technique appears rapid and efficacious in the case of visible intracoronary thrombus.

Our purpose is to review the different mechanisms involved in the vascular response following percutaneous coronary revascularization, and their role in thrombotic complications and in restenosis. Specificities related to the various techniques of percutaneous coronary revascularization (balloon angioplasty, coronary stents, brachytherapy, drug eluting stents) are discussed.

Systolic heart failure is mainly due to a decreased left ventricular systolic function after myocardial infarction. Despite significant improvements in medical management of heart failure, the prognostic remains poor, with a 5-years survival only of 30%.Current treatments are only able to delay progression toward end-stage heart failure. Heart transplantation is accessible only for selected patients. Thus, in the context of post-myocardial infarction, cell therapy appears to be a new original technique, available for the majority of patients and potentially non-invasive.After promising results in experimental models, a phase I clinical trial has been conducted in France, showing the feasibility of intracardiac autologous skeletal myoblast implantation. Other studies in Europe and USA are currently testing a variety of cells and delivery systems. A phase II-trial will begin in France.

Introduction En therapie cellulaire du diabete, le rapport benefices de l'optimisation glycemique/risques cardiovasculaires du traitement immuno-suppresseur reste mal connu. Notre but etait de determiner l'evolution des complications macroangiopathiques 5 ans apres greffe d'ilots. Patients et methodes 21 des 36 patients greffes suivis prospectivement a Lille, avaient au moins 5 ans de recul et ont ete inclus. Leurs caracteristiques initiales etaient les suivantes : diabete insulinoprive depuis 28±9 ans; âge : 43±7 ans; IMC : 24±2kg/m 2 ; 8/21 greffes renales 23±10 mois auparavant; immunosuppression : Edmonton. Ces 21 patients ont beneficie avant greffe puis annuellement d'un depistage systematique des complications macroangiopathiques. L'analyse a ete faite en intention de traiter. Trois « ilots-seuls » et 2 « ilots-apres-rein » avaient perdu leur fonction C-peptidique a 5 ans. Neuf patients etaient insulino-independants dont 8 avaient une HbA1c 6,5 % (38 % avec les 2 criteres). La moyenne d'HBA1c etait de 8,1±1,0 % avant vs. 7,1±1,0 % apres greffe (p Resultats Les moyennes de cholesterol total (1,86±0,37vs.1,76±0,36g/l), HDL (0,71±0,18vs.0,73±0,27g/l), LDL (0,98±0,32vs.0,88±0,30g/l), triglycerides (0,79±0,33vs.0,83±0,34g/l) et pression arterielle moyenne (124±17/72±7 vs. 127±13/72±8mmHg) ne differaient pas avant vs. apres greffe (patients traites par : statine : 28 % vs.65 %; 1 antihypertenseur : 38 % vs.75 %). Trois patients dont 2 « ilots-apres-rein » avaient des complications macroangiopathiques avant greffe (2 pontages femoraux dont 1 avec amputation, 1 stent coronarien). Le nombre d'evenements cardiovasculaires clinique pendant ces 5 ans etait nul. Avant vs. apres greffe, le depistage etait anormal dans : echodoppler carotidien : 37 % vs 52 % des cas; membres inferieurs : 36 % vs 85 % (22 vs 25 % hors mediacalcose simple), recherche d'ischemie : 14 % (avec coronarographie normale hormis 1 calcification) vs 19 % (dont 3 confirmees en coronarographie, stentees). Conclusion La greffe n'a induit aucun evenement cardiovasculaire aigu clinique chez ces 21 patients atteints d'un diabete ancien, dont 8 greffes renaux au prix du depistage systematique et du traitement des complications macroangiopathiques silencieuses.

The identification of coronary thrombosis in humans has important prognostic and therapeutic implications. Its recognition calls for invasive techniques: angioscopy, coronarography, and endocoronary ultrasound. Angioscopy allows visualisation of the arterial surface and the detection of thrombus with great sensitivity. Its presence is exclusive to the acute coronary syndromes: 67% in unstable angina, and 75% in myocardial infarction, although it is only present in 27% of cases of stable angina. The relative complexity of its use has led to the abandonment of this technique. Coronarography allows an indirect approach to coronary thrombosis. Certain aspects are evocative such as: intraluminal filling defect, complete occlusion with upstream convexity, ulceration and eccentric type 2 Ambrose classification plaques. As a function of the clinical presentation, the coronarographic views allow a good specificity for the diagnosis of thrombus. The sensitivity is weak, however, compared to angioscopy. Endocoronary ultrasound does not allow identification of fresh thrombus which is not echogenic and does not allow differentiation between older thrombus and lipid plaque. In the future, magnetic resonance imaging could prove interesting in the detection of recent thrombus.

Though the efficacy of intravascular gene transfer has been demonstrated in native vessels following acute injury, this methodology has not been validated in complex models of vascular injury that more closely mimic clinical angioplasty procedures. Previous studies have shown that Gax gene overexpression modulates the injury-induced remodeling of the vessel in rat carotid and normal rabbit iliac arteries. Here, we evaluated the effect of the Gax gene delivery in atheromatous stented vessels. Rabbits were fed 120 g daily of 1% cholesterol diet for 3 weeks. At 1 week they underwent initial injury on the external iliac artery, then balloon angioplasty was performed at 3 weeks at the same site with a 2.5 mm diameter channel balloon catheter (three times 1 min at 6 atm). Either saline (n = 4) or the control viral construct Ad-CMVluc (5 x 109 p.f.u.) (n = 5) or Ad-CMVGax (5 x 10(9) p.f.u.) (n = 4) was delivered with a poloxamer mixture via a channel balloon (6 atm, 30 min), and a 15 mm long Palmaz-Schatz stent (PS154) was then deployed at the site (1 min, 8 atm). Arteries were analyzed 1 month later. At 1 month, the Ad-CMVGax treated arteries exhibited a lower maximal intimal area (1. 15+/-0.1 mm2) than saline (1.87+/-0.15 mm2, P = 0.007) or Ad-CMVluc-treated vessels (1.98+/-0.31 mm2, P = 0.04). Likewise Ad-CMVGax-treated vessels displayed a lower maximal percentage cross-sectional area narrowing (35.1+/-3.5%) than saline (65.3+/-9.4%, P = 0.01) or Ad-CMVluc-treated vessels (62.7+/-6.7%, P = 0.02). Angiographic analysis revealed larger minimal lumen diameter in Ad-CMVGax treated arteries (2.0+/-0.1 mm) than saline (1.14+/-0.36 mm, P = 0.06) or Ad-CMVluc-treated vessels (1.23+/-0.25 mm, P = 0.02). Overexpression of the Gax gene inhibits neointimal hyperplasia and lumen loss in atheromatous stented rabbit iliac arteries.

In a multicenter, randomized trial, systematic stenting using the Wiktor stent was compared to conventional balloon angioplasty with provisional stenting for the treatment of acute myocardial infarction (AMI).Primary angioplasty in AMI is limited by in-hospital recurrent ischemia and a high restenosis rate.A total of 211 patients with AMI12 h from symptom onset, with an occluded native coronary artery, were randomly assigned to systematic stenting (n = 101) or balloon angioplasty (n = 110). The primary end point was the binary six-month restenosis rate determined by core laboratory quantitative angiographic analysis.Angiographic success (Thrombolysis in Myocardial Infarction [TIMI] flow grade 3 and residual diameter stenosis50%) was achieved in 86% of the patients in the stent group and in 82.7% of those in the balloon angioplasty group (p = 0.5). Compared with the 3% cross-over in the stent group, cross-over to stenting was required in 36.4% of patients in the balloon angioplasty group (p = 0.0001). Six-month binary restenosis (or = 50% residual stenosis) rates were 25.3% in the stent group and 39.6% in the balloon angioplasty group (p = 0.04). At six months, the event-free survival rates were 81.2% in the stent group and 72.7% in the balloon angioplasty group (p = 0.14), and the repeat revascularization rates were 16.8% and 26.4%, respectively (p = 0.1). At one year, the event-free survival rates were 80.2% in the stent group and 71.8% in the balloon angioplasty group (p = 0.16), and the repeat revascularization rates were 17.8% and 28.2%, respectively (p = 0.1).In the setting of primary angioplasty for AMI, as compared with a strategy of conventional balloon angioplasty, systematic stenting using the Wiktor stent results in lower rates of angiographic restenosis.

Objective: Growth regulatory properties of nitric oxide (NO) in cultured endothelial cells is controversial. The aim of our study was to investigate the effect of l-arginine, the endogenous NO precursor, and l-NAME, an inhibitor of NO synthase on the reendothelialization process after angioplasty. Methods: Fifty-five New Zealand White rabbits underwent denudation of the left iliac artery. After injury the rabbits were randomized in three groups: l-arginine 2.25% (l-arginine, n =19); N G-nitro-l-arginine methyl ester 15 mg/kg/day (l-NAME, n =19); and placebo (controls, n =17). Treatment was solubilized in drinking water. Reendothelialization was evaluated at 4 weeks by macroscopic evaluation of Evans blue staining and endothelial-specific immunostaining (CD-31) on cross sections. Intimal hyperplasia was evaluated by morphometric analysis. Results: Despite a significant increase in plasma arginine ( P =0.001) and a reduction in intimal hyperplasia ( P =0.003) with l-arginine, neither agent had a significant effect on reendothelialization at 4 weeks (controls=36±4%, l-arginine=43±3%, l-NAME=33±4%; NS). Conclusion: These results suggest that, in spite of previously demonstrated effects on neointimal hyperplasia, the NO pathway does not influence the regrowth of macrovascular endothelial cells in vivo.

The next five years we will be swamped with stem cell papers, presentations, working groups and task forces. Maybe there is already a society for stem cells and cardiovascular therapy. If there is something potentially interesting on the horizon, we all start running instead of thinking.1 There are numerous stem cell variants, sources and ways to apply them. There are also many diseases that need a better and more aetiological treatment then just pharmacology. Clearly, the stem cell field and the expectations provide all the ingredients for a hype. So let us focus on the horizon, forget the obstacles and enjoy the hype for as long as it lasts.2-8

Background —Dual therapy with ticlopidine and aspirin has been shown to be as effective as or more effective than conventional anticoagulation in patients with an optimal result after implantation of intracoronary metallic stents. However, the safety and efficacy of antiplatelet therapy alone in an unselected population has not been evaluated. Methods —Patients were randomized to conventional anticoagulation or to treatment with antiplatelet therapy alone. Indications for stenting were classified as elective (decided before the procedure) or unplanned (to salvage failed angioplasty or to optimize the results of balloon angioplasty). After stenting, patients received aspirin and either ticlopidine or conventional anticoagulation (heparin or oral anticoagulant). The primary end point was the occurrence of bleeding or peripheral vascular complications; secondary end points were cardiac events (death, infarction, or stent occlusion) and duration of hospitalization. Results —In 13 centers, 236 patients were randomized to anticoagulation and 249 to antiplatelet therapy. Stenting was elective in 58% of patients and unplanned in 42%. Stent implantation was successfully achieved in 99% of patients. A primary end point occurred in 33 patients (13.5%) in the antiplatelet group and 48 patients (21%) in the anticoagulation group (odds ratio=0.6 [95% CI 0.36 to 0.98], P =0.03). Major cardiac-related events in electively stented patients were less common (odds ratio=0.23 [95% CI 0.05 to 0.91], P =0.01) in the antiplatelet group (3 of 123, 2.4%) than the anticoagulation group (11 of 111, 9.9%). Hospital stay was significantly shorter in the antiplatelet group (4.3±3.6 versus 6.4±3.7 days, P =0.0001). Conclusions —Antiplatelet therapy after coronary stenting significantly reduced rates of bleeding and subacute stent occlusion compared with conventional anticoagulation.

Background —Scatter factor/hepatocyte growth factor (SF/HGF) is a pleiotropic growth factor that stimulates proliferation and migration of endothelial cells (ECs) via the c- Met receptor, present on ECs as well as other cell types, including smooth muscle cells (SMCs). We studied the effects of recombinant human (rh) SF/HGF in vitro and in vivo in a rabbit model of hindlimb ischemia. We further compared these effects with those of recombinant human vascular endothelial growth factor (rhVEGF 165 ), an EC–specific mitogen. Methods and Results —In vitro, rhSF/HGF and rhVEGF 165 exhibited similar effects on proliferation and migration of ECs. When both cytokines were administered together, the result was an additive effect on EC proliferation and a synergistic effect on EC migration. Application of rhSF/HGF to cultures of human SMCs resulted in the induction of VEGF mRNA and protein. In vivo, administration of rhSF/HGF (500 μg×3) was associated with significant improvements in collateral formation ( P P P 165 administered according to the same protocol ( P 165 . Conclusions —The pleiotropic effects of certain growth factors may potentiate angiogenesis via a combination of direct effects on EC proliferation and migration and indirect effects that result in the generation of other potent EC mitogens from non-EC populations. The synergistic effects demonstrated when SF/HGF and VEGF are administered together in vitro may be reproduced in vivo by SF/HGF-induced upregulation of VEGF in vascular SMCs.

Objectives: The expression of gax , an anti-proliferative homeobox gene, is rapidly downregulated in vascular smooth muscle cells (VSMCs) following arterial injury. Here we performed percutaneous adenovirus-mediated gene transfer into the iliac arteries of normal rabbits using a channel balloon catheter to assess the effects of gax overexpression on neointima formation, lumen diameter, reendothelialization and functional vasomotion. Methods: A channel balloon catheter was used to perform both the arterial injury and local gene delivery. In each animal both iliac arteries were randomly assigned to receive either an adenovirus expressing the gax gene (Ad-Gax) or the β-galactosidase gene (Ad-βgal). In a second group of animals arteries were randomly assigned to receive either Ad-βgal or saline. Results: At one month, angiography revealed 36% less luminal narrowing in the Ad-Gax-treated arteries relative to the Ad-βgal-treated control arteries. Histological analysis revealed that the intimal/medial ratio (I/M) was reduced by 56% in the Ad-Gax group. Endothelium-dependent vasomotion was not affected by the gax gene transfer. In the second group, no statistically significant differences were found between the saline and the Ad-βgal-treated vessels for any of these parameters. Conclusions: Percutaneous adenovirus delivery of the gax gene to rabbit iliac arteries following endothelial denudation and vessel wall injury reduces neointimal hyperplasia and luminal stenosis, but does not affect endothelium-dependent vasomotion. This study demonstrates that a VSMC transcription factor can potentially be utilized for the development of a molecular therapy for vascular disorders.

This study sought to determine whether pravastatin affects clinical or angiographic restenosis after coronary balloon angioplasty.Experimental data and preliminary clinical studies suggest that lipid-lowering drugs might have a beneficial effect on restenosis after coronary angioplasty.In a multicenter, randomized, double-blind trial, 695 patients were randomized to receive pravastatin (40 mg/day) or placebo for 6 months after successful balloon angioplasty. All patients received aspirin (100 mg/day). The primary angiographic end point was minimal lumen diameter (MLD) at follow-up, assessed by quantitative coronary angiography. A sample size of 313 patients per group was required to demonstrate a difference of 0.13 mm in MLD between groups (allowing for a two-tailed alpha error of 0.05 and a beta error of 0.20). To allow for incomplete angiographic follow-up (estimated lost to follow-up rate of 10%), 690 randomized patients were required. Secondary end points were angiographic restenosis rate (restenosis assessed as a categoric variable,50% stenosis) and clinical events (death, myocardial infarction, target vessel revascularization).At baseline, clinical, demographic, angiographic and lipid variables did not differ significantly between groups. In patients treated with pravastatin, there was a significant reduction in total and low density lipoprotein cholesterol and triglyceride levels and a significant increase in high density lipoprotein cholesterol levels. At follow-up the MLD (mean +/- SD) was 1.47 +/- 0.62 mm in the placebo group and 1.54 +/- 0.66 mm in the pravastatin group (p = 0.21). Similarly, late loss and net gain did not differ significantly between groups. The restenosis rate (recurrence50% stenosis) was 43.8% in the placebo group and 39.2% in the pravastatin group (p = 0.26). Clinical restenosis did not differ significantly between groups.Although pravastatin has documented efficacy in reducing clinical events and angiographic disease progression in patients with coronary atherosclerosis, this study shows that it has no effect on angiographic outcome at the target site 6 months after coronary angioplasty.

The carbohydrate moieties of glycoprotein hormones or growth factor molecules may have a variety of effects that impact biological potency. Vascular endothelial growth factor (VEGF), also known as vascular permeability factor (VPF), is a 45 kD heparin-binding, endothelial cell (EC) specific mitogen with a putative N-linked glycosylation site. Recent studies have shown that VEGF/VPF may successfully augment collateral development in animal models of myocardial and hindlimb ischemia. The extent to which glycosylation of the 75 asparagine site affects the angiogenic properties of VEGF/VPF has not been studied in vivo. Specifically unaddressed to date is the concern that nonglycosylated VEGF/VPF may be less stable, and therefore characterized by a shorter half-life, reducing its utility for therapeutic angiogenesis. Accordingly, the purpose of this study was to investigate the extent to which posttranslational modification, specifically glycosylation, mofies the angiogenic properties of VEGF/VPF in vivo. Glycosylated (g+) recombinant human VEGF165 was purified from media conditioned by Chinese hamster ovary (CHO) cells. Nonglycosylated (g-) VEGF165 was expressed, purified and refolded from E. coli. The purity of both materials was assessed by silver-stained SDS/PAGE and characterized by the presence of a single amino terminal sequence as indicated by Edman degradation. Tryptic mapping by reverse-phase HPLC confirmed that the potential glycosylation site at 75 asparagine was occupied by N-linked carbohydrate for the Chinese hamster ovary-derived VEGF/VPF, but not for E. coli-derived VEGF/VPF. The mitogenic effects of Chinese hamster ovary-derived (g+) VEGF165 and E. coli-derived (g-) VEGF165 wre studied in vitro using microvascular EC. At concentrations of VEGF/VPF ranging from 10(-4) to 10(2) nM, both produced similar concentration-dependent effects on EC proliferation. For in vivo studies, (g-) (n = 8) and (g+) (n = 8) formulations of VEGF/VPF were administered to New Zealand white rabbits with unilateral hindlimb ischemia. For (g-) versus (g+) VEGF/VPF-treated groups, respectively, calf blood pressure ratio was 0.40 +/- 0.04 versus 0.37 +/- 0.04; angiographic score (of collateral vessels) was 0.37 +/- 0.04 versus 0.35 +/- 0.04; capillary density (capillaries/mm2) at necropsy was 246.9 +/- 21.5 versus 253.9 +/- 18.8; and tissue perfusion (colored microspheres) was 92.8 +/- 5.5 versus 90.30 +/- 13.47 (all p = ns). Moreover, intravascular Doppler-based analyses of resting, maximum, and endothelium-dependent flow was similar for (g-) and (g+) VEGF/VPF. These in vitro and in vivo findings establish that the potential for VEGF/VPF to stimulate therapeutic angiogenesis persists unaltered in the nonglycosylated state.

Angiotensin-converting enzyme (ACE) inhibitors reduce intimal hyperplasia after balloon injury. A role for nitric oxide (NO) has been suggested in this effect. Because recent data suggest that NO may modulate some features of endothelial cells and because endothelial cells are involved in the control of intimal hyperplasia, we investigated the role of NO synthesis in the effect of an ACE inhibitor, perindopril, on neoendothelial dysfunction and intimal hyperplasia in a rabbit model of unilateral iliac balloon injury. New Zealand White male rabbits received placebo, perindopril, or cotreatment with perindopril and NG-nitro-L-arginine methyl ester (L-NAME) and were evaluated 4 wk after the injury. Fifteen rabbits (5 in each group) were used to assess in vitro vasoreactivity and twenty-four (8 in each group) for morphometric analysis. In injured vessels, neoendothelium-dependent relaxation in ACE inhibitor-treated animals was improved compared with placebo (P < 0.05) and restored to the level of noninjured vessels (NS). The improvement observed with ACE inhibitor was abolished by cotreatment with L-NAME (P < 0.05). In the same vessels, no effect was observed on neoendothelium-independent vasoreactivity. The improved neoendothelial dysfunction with ACE inhibitor was associated with a 66% reduction in intimal thickening (P < 0.01). The effect was also reversed by cotreatment with L-NAME (P < 0.01). In noninjured vessels, treatment did not alter vasoreactivity or morphology of the vessel wall. These results suggest that NO synthesis may play a key role in the improvement of vascular function seen with ACE inhibitor in balloon-injured vessels.

Acute coronary occlusion as a consequence of dissection or thrombosis occurs in 2 to 11% of patients treated with percutaneous transluminal coronary angioplasty (PTCA), and continues to be the principal cause of early morbidity and mortality. In this study the experience of one center is presented with the application of two types of stents, Wiktor (Medtronic Inc.) or Gianturco-Roubin (Cook Inc.) for bailout of acute or threatening coronary occlusion that persisted after treatment with prolonged balloon inflation. All patients received a complete anticoagulation scheme with heparin, dextran, dipyridamole, aspirin and coumadin. From January to November 1993, 26 patients with 27 prosthesis were included. There were 21 men and 5 women with mean age of 58 years (range 36 to 73). The indications for stenting were: total occlusion in five (19%) threatening occlusion in 13 (50%) and severe persistent dissection in eight (31%). Initial implantation success was 93% (25/27). Procedure related clinical complications were death in one patient, bypass surgery in two (8%) and myocardial infarction in four (15%). Acute stent thrombosis occurred in three cases and subacute in one (11 and 4% respectively). Three patients, had non-fatal bleeding complications. Final clinical success without myocardial infarction, bypass surgery or death was 77%. In conclusion, coronary stenting for bailout of acute or threatening coronary occlusion after PTCA is a good alternative to emergency surgery. New antithrombotic strategies and better anticoagulation schemes may improve further this procedure.

The aim was to examine the effects of aldosterone and of an aldosterone antagonist, spironolactone, on neointimal thickening in a rabbit model of balloon injury.Eighteen rabbits underwent aortic and iliac balloon injury and were randomised to subcutaneous infusion of aldosterone (70 micrograms.kg-1.d-1) or vehicle solution for 28 d. Eighteen other rabbits were randomised to receive daily subcutaneous injections of spironolactone (50 mg.kg-1.d-1) or of vehicle for 7 d before injury and for 28 d after the procedure. All animals were then killed just after measurement of plasma renin activity and of arterial pressure. Vessels were fixed and five cross sections were analysed per rabbit (three aortic; two from iliac artery). Mean values of neointimal area and of the neointimal area/medial area ratio were calculated.Aldosterone treatment was associated with a decrease in renin activity and a non-significant increase in mean arterial pressure. Aldosterone significantly augmented the neointimal thickening in the iliac artery [0.42(SEM 0.07) v 0.24(0.03) mm2, P0.05] but not in the aorta [0.63(0.08) v 0.59(0.12) mm2, NS]. Spironolactone significantly inhibited intimal thickening, both in the iliac artery [0.09(0.02) v 0.29(0.01) mm2, P0.001] and in the aorta [0.31(0.03) v 0.59(0.06) mm2, P0.001]. Spironolactone administration was associated with an increase in renin activity and a decrease in mean arterial blood pressure.Aldosterone administration enhances neointimal thickening after injury and spironolactone, an aldosterone antagonist, is a potent inhibitor of neointimal thickening in the same model. This suggests a role for aldosterone in the pathophysiology of neointimal proliferation after balloon injury and for aldosterone antagonists in its prevention.

BACKGROUND Angiopeptin, a synthetic cyclic octapeptide analogue of somatostatin, reduces neointimal hyperplasia after balloon denudation of rabbit aorta if administered before injury. The aim of this study was to analyze the effect of angiopeptin pretreatment on the level of expression of the c-fos and c-jun protooncogenes, early markers of smooth muscle cell proliferation, after balloon denudation of rabbit aorta. METHODS AND RESULTS For histological analysis of the effect of angiopeptin on neointimal thickening after aortic balloon denudation, rabbits were randomized into three groups: group 1 (controls), twice-daily injections of saline begun 24 hours before balloon denudation (n = 9); group 2, twice-daily injections of angiopeptin 10 micrograms/kg begun 24 hours before balloon denudation (n = 9); and group 3, twice-daily injections of angiopeptin 10 micrograms/kg begun 1 hour after balloon denudation (n = 7). The degree of neointimal thickening 28 days after balloon denudation was significantly less in group 2 than in group 1 (neointimal area: group 1, 0.59 +/- 0.11 mm2; group 2, 0.22 +/- 0.05 mm2; P < .05. Neointima/media: group 1, 0.85 +/- 0.17; group 2, 0.23 +/- 0.05; P < .05). When angiopeptin was started 1 hour after denudation (group 3), however, the neointimal area (0.52 +/- 0.09 mm2) and the neointima/media ratio (0.76 +/- 0.10) were not statistically different from the control group. For analysis of protooncogene induction, rabbits received twice-daily subcutaneous injections of saline (n = 7), angiopeptin 10 micrograms/kg (n = 8), or angiopeptin 100 micrograms/kg (n = 4) begun 24 hours before balloon denudation. The animals were killed 30 minutes after balloon denudation, and total aortic RNA was hybridized with fos and jun probes. Expression of c-fos and c-jun was detected 30 minutes after injury; angiopeptin pretreatment at 20 micrograms.kg-1.d-1 induced a 41% reduction in c-fos expression and a 42% reduction in c-jun expression compared with control animals. The inhibitory effect at the higher dose of angiopeptin was similar. CONCLUSIONS Our results show that the inhibitory effect of angiopeptin on neointimal thickening is related to events that occur very early after injury and suggest that the inhibition of smooth muscle cell activation may be responsible, at least in part, for this effect.

Contexte Le systeme renine-angiotensine-aldosterone constitue une cible therapeutique majeure dans le traitement de la maladie coronarienne. Des travaux recents suggerent l’interet pronostique de l’aldosterone dans les syndromes coronariens aigus (SCA). Nous avons etudie la valeur pronostique potentielle de l’aldosterone dans une population de coronariens plus large : ceux beneficiant d’une angioplastie coronaire programmee. Methodes De juin 2000 a Septembre 2001, nous avons inclus les patients hospitalises au CHRU de Lille beneficiant d’une angioplastie coronaire non urgente. Des prelevements sanguins etaient realises durant la coronarographie pour dosage de l’aldosterone plasmatique et le suivi etait d’au moins 12 mois, le critere de jugement etant la mortalite cardiovasculaire. Resultats 807 patients ont ete inclus, âges en moyenne de 61 ans, essentiellement des hommes (78 %) fumeurs (71 %) hypertendus (53 %). 32 % etaient diabetiques, la FEVG moyenne etait de 58 ± 15 % et 93 % des patients ont recu une endoprothese. 50 % des patients etaient des coronariens stables. Le taux moyen d’aldosterone etait de 25 (13-5) pg/mL. Les facteurs associes a une augmentation de l’aldosterone etaient l’IMC (p = 0.003), une classe NYHA > 1 (p = 0.0001) et une elevation de la troponine (p = 0.01). Inversement un âge eleve (p = 0.0001), une fonction renale normale (p = 0.01) et la prise de betabloquants (p = 0.01) etaient associes a une diminution de l’aldosterone. Le suivi moyen etait de 14.9 mois et il y a eu 40 deces d’origine cardiovasculaire. Les facteurs predictifs independants de mortalite cardiovasculaire en analyse multivariee etaient un âge eleve (RR = 1.42, p = 0.04), la presence d’un diabete (RR = 2.20, p = 0.04), une fraction d’ejection alteree (RR = 1.58, p = 0.001), la survenue recente d’un SCA (RR = 3.23, p = 0.02), une CRP elevee (RR = 2.59, p = 0.004) et un taux d’aldosterone eleve (RR = 3.48, p = 0.004). Conclusion L’aldosterone plasmatique semble etre un facteur pronostique independant chez les patients beneficiant d’une angioplastie coronaire non urgente et son utilisation permettrait d’ameliorer la stratification du risque cardiovasculaire. Cela pourrait n’etre que le reflet de l’activation generale du systeme neurohormonal mais l’aldosterone induit divers effets extrarenaux (developpement d’une dysfonction endotheliale, d’une fibrose vasculaire et myocardique, effets pro-thrombotiques et proinflammatoires) qui en font un marqueur de risque cardiovasculaire tres seduisant. Download : Download full-size image

Le Tissue Factor Pathway Inhibitor-2 (TFPI-2) est un inhibiteur de sérineprotéase de type Kunitz exprimé dans la plaque d’athérosclérose. Le TFPI-2 y présenterait un effet athéroprotecteur en neutralisant les métalloprotéases matricielles (MMP). Le promoteur du gène du TFPI-2 présente plusieurs ilôts CpG dont l’hyperméthylation est associée à une diminution de son expression. Cette régulation épigénétique de l’expression du TFPI-2 a été mise en évidence dans la cancer mais n’a pas encore été étudiée dans l’athérosclérose. L’objectif de ce travail a été d’étudier la méthylation du gène du TFPI-2 dans l’athérosclérose carotidienne avec ses conséquences sur l’expression du transcrit TFPI-2. Le statut de méthylation du gène du TFPI-2 a également été confronté aux données cliniques des patients et à la morphologie des plaques.La méthylation de 18 motifs CpG du promoteur du TFPI-2 a été étudiée par Methylation Specific PCR (MSP) et pyroséquençage (PyroMark MD, Biotage) dans 59 plaques d’athérosclérose carotidienne et 26 artères mammaires contrôle.Seize plaques (27 %) étaient retrouvées méthylées par MSP (groupe MSP+) alors qu’aucune artère mammaire ne l’était. Le pyroséquençage a confirmé que les plaques MSP+ présentaient des niveaux de méthylation significativement plus élevés que les plaques MSP- mais aussi que les artères (p = 0,03 et 0,01, respectivement). Le niveau de transcrit TFPI-2 était significativement plus bas dans les plaques méthylées que dans les non méthylées et que dans les artères (p=0,04 et < 0,0001, respectivement). Les facteurs de risque cardiovasculaire, les accidents ischémiques cérébraux mais également l’activité MMP-9 des plaques n’étaient pas différents entre les échantillons méthylés et non méthylés. Cependant, les plaques méthylées présentaient des infiltrats lipidiques et macrophagiques réduits par rapports aux plaques non méthylées.Cette étude démontre pour la première fois que l’expression du TFPI-2 peut être régulée par hyperméthylation de son gène dans l’athérosclérose. L’hyperméthylation du gène du TFPI-2 est associée à une diminution de l’expression de son transcrit mais également à des plaques morphologiquement moins sévères. L’impact de la méthylation du TFPI-2 sur la régulation des MMP et sur la fragilité de la plaque d’athérosclérose reste à déterminer.

Objectives : Angiotensin converting enzyme (ACE) inhibitors reduce neointimal hyperplasia after balloon denudation, but the mechanisms are not completely understood. It has been demonstrated that nuclear oncogenes are induced in the vascular wall in the hours immediately after injury, and that the same genes are induced by angiotensin II in vascular smooth muscle cells. It has therefore been suggested that the effects of ACE inhibitors on the response of the vessel wall could be mediated by an inhibition of proto-oncogene expression. Methods and results : Sixteen New Zealand White rabbits were randomly assigned for histologic analysis to receive placebo (n=9) or 1 mg/kg per day perindopril (n=7). After treatment for 7 days balloon aortic injury was performed. The treatment was continued and the rabbits were killed 28 days after injury. In the perindopril group the neointimal cross-sectional area was significantly smaller than in the control group. Six untreated rabbits were used to assess the time course of proto-oncogene expression in the aortic wall after injury in the present model. After extraction, total aortic RNA was hybridized with myc, fos and jun probes. Based on the results, the effects of ACE inhibition on proto-oncogene expression were tested 1 h after balloon denudation. Accordingly, 24 rabbits were randomly assigned to pretreatment for 7 days with placebo or with 1 or 10 mg/kg per day perindopril (n = 8, for each group) and were killed 1 h after injury. Expression of c-myc was not altered by pretreatment. However, 1 mg/kg per day perindopril induced significant reductions of 50% in c-jun and 45% in c-fos expression compared with control. No additional effect was obtained with the higher dose. Conclusion : The effect of ACE inhibition on intimal hyperplasia is associated with a reduction in early cellular events such as c-fos and c-jun expression. These results suggest that potent ACE inhibition at the time of vascular injury may be required to limit the hyperplastic response of the vessel wall.

Background Patients with critical limb ischaemia (CLI) unsuitable for revascularisation have a high rate of amputation and mortality (30% and 25% at 1 year, respectively). Localised gene therapy using plasmid DNA encoding acidic fibroblast growth factor (NV1FGF, riferminogene pecaplasmid) has showed an increased amputation-free survival in a phase II trial. This article provides the rationale, design and baseline characteristics of CLI patients enrolled in the pivotal phase III trial (EFC6145/TAMARIS). Methods An international, double-blind, placebo-controlled, randomised study composed of 525 CLI patients recruited from 170 sites worldwide who were unsuitable for revascularisation and had non-healing skin lesions was carried out to evaluate the potential benefit of repeated intramuscular administration of NV1FGF. Randomisation was stratified by country and by diabetic status. Results The mean age of the study cohort was 70 ± 10 years, and included 70% males and 53% diabetic patients. Fifty-four percent of the patients had previous lower-extremity revascularisation and 22% had previous minor amputation of the index leg. In 94% of the patients, the index leg had distal occlusive disease affecting arteries below the knee. Statins were prescribed for 54% of the patients, and anti-platelet drugs for 80%. Variation in region of origin resulted in only minor demographic imbalance. Similarly, while diabetic status was associated with a frequent history of coronary artery disease, it had little impact on limb haemodynamics and vascular lesions. Conclusions Clinical characteristics and vascular anatomy of CLI patients with ischaemic skin lesions who were unsuitable for revascularisation therapy show little variations by region of origin and diabetic status. The findings from this large CLI cohort will contribute to our understanding of this disease process. This study is registered with ClinicalTrials.gov , number NCT00566657.

In animal models, a hypercholesterolemic diet induces areas of deendothelialization and impairs endothelium-dependent vasorelaxation. Angiogenic growth factors increase endothelial cell growth in vivo. This study was thus designed to test the hypothesis that chronic administration of basic fibroblast growth factor (bFGF) in hypercholesterolemic rabbits might restore normal physiological responses to endothelium-dependent agonists. After feeding on a 2% cholesterol diet for 6 wk, 14 New Zealand White rabbits received twice-weekly intravenous boluses of either 2.5 microg bFGF (hypercholesterolemic bFGF group, n = 6) or placebo (hypercholesterolemic control group, n = 8) for 3 wk and were killed for assessment of in vitro vasoreactivity and for histological analysis. Six animals fed with standard rabbit diet were used to assess normal responses. Endothelium-dependent responses to acetylcholine and to the calcium ionophore A-23187 were reduced in the hypercholesterolemic control group compared with normal rabbits. Hypercholesterolemic animals treated with bFGF had significantly better endothelium-dependent responses than hypercholesterolemic rabbits not treated with bFGF. Endothelium-independent responses did not differ significantly among the three groups. A similar degree of plaque formation was observed in control- and bFGF-treated hypercholesterolemic rabbits. These results show that, in this model of atherosclerosis, bFGF has a highly beneficial effect on the functional responses of atherosclerotic vessels and does not have a deleterious effect on the degree of plaque formation.